Molecular mechanisms of apoptosis in hepatocellular carcinoma cells induced by ethanol extracts of Solanum lyratum Thumb through the mitochondrial pathway

Table 1 Reverse transcription polymerase chain reaction primers

Gene name	Primer sequence	Product length, bp
Fas	forward: 5′-GAATGCAAGGGACTGATAGC-3′	414
	Reverse: 5′-TGGTTCGTGTGCAAGGCTC-3′
FasL	forward: 5′-GGAATGGGAAGACACATATGGAACTGC-3′	239
	Reverse: 5′-CATATCTGGCCAGTAGTGCAGTAATT-3′
Caspase-3	forward: 5′-GCTATTGTGAGGCGGTTGT-3′	270
	Reverse: 5′-CGTTTGGAGTCCCTTTGT-3′
Caspase-8	forward: 5′-TCTGGAGCATCTGCTGTCTG-3′	427
	Reverse: 5′-TTGACGTCTGTGGTCCGTCC-3′
p53	forward: 5′-CTTTCAACTCTGTCTCCTTC-3′	180
	Reverse: 5′-TGGGCAACCAGCCCTGTCGT-3′
Bcl-2	forward: 5′-CAGCTGCACCTGACGCCCTT-3′	231
	Reverse: 5′-GCCTCCGTTATCCTGGATCC-3′
β-actin	forward: 5′-ACACTGTGCCCATCTACGAGG-3′	621
	Reverse: 5′-AGGGGCCGGACTCGTCATACT-3′

Table 2 Comparison of the inhibition rates of proliferation of hepatocellular carcinoma SMMC-7721 cells in the groups with different Solanum lyratum Thumb concentrations, ^-x ± s

Group	Concentration, mg/L	n	OD value	Inhibition rate, %
Three ST groups with different concentrations	2.5	12	1.516 ± 0.205	18.22 ± 2.32
	5	12	1.242 ± 0.236	36.56 ± 2.51
	10	12	0.815 ± 0.276	55.98 ± 3.12
Negative control group	0	12	1.842 ± 0.183	-
Positive control group	2.5	12	1.135 ± 0.172	38.60 ± 1.78

The comparison between ST groups and the negative control group shows that P < 0.05, while that of ST groups and the positive control group indicates that P > 0.05. ST: Solanum lyratum Thumb.

Table 3 Comparison of apoptosis rates of hepatocellular carcinoma SMMC-7721 cells in different groups, ^-x ± s

Group	Concentration, C/mg•L-1	Apoptosis rate of SMMC-7721 cells, %
Three ST groups with different concentrations	2.5	10.75 ± 2.51
	5	17.31 ± 2.33
	10	30.21 ± 3.62
Negative control group	0	3.75 ± 1.23
Positive control group	2.5	17.36 ± 1.62

The comparison of the ST groups and the negative control group shows that P < 0.05, while that of ST groups with the positive control group reveals that P > 0.05. ST: Solanum lyratum Thumb.

Table 4 mRNA expressions of genes relating to hepatocellular carcinoma SMMC-7721 cells in each group

Group	Concentration, C/mg•L-1	mRNA expression of genes
		Fas	FasL	Caspase-8	Caspase-3	p53	Bcl-2	β-actin
Three ST groups with different concentrations	2.5	52.47 ± 0.10	58.25 ± 0.51	30.57 ± 0.10	22.15 ± 0.02	34.17 ± 0.01	36.28 ± 0.14	17.53 ± 0.14
	5	52.74 ± 0.08	56.81 ± 0.30	30.76 ± 0.03	22.33 ± 0.09	34.45 ± 0.20	35.40 ± 0.51	18.15 ± 0.41
	10	53.41 ± 0.64	55.94 ± 0.80	31.30 ± 0.24	23.01 ± 0.47	35.11 ± 0.69	33.54 ± 0.51	19.10 ± 0.91
Negative control group	0	52.32 ± 0.04	59.13 ± 0.07	30.44 ± 0.06	22.05 ± 0.03	34.11 ± 0.02	37.59 ± 0.33	17.30 ± 0.17
Positive control group	2.5	52.79 ± 0.11	56.69 ± 0.34	30.77 ± 0.03	22.35 ± 0.12	34.43 ± 0.20	35.53 ± 0.35	18.18 ± 0.41

ST: Solanum lyratum Thumb.

Table 5 Differences in mRNA expressions of genes relating to hepatocellular carcinoma SMMC-7721 cells in each group

Group	Concentration, C/mg•L-1	2-ΔΔCT
		Fas	FasL	Caspase-8	Caspase-3	p53	Bcl-2
Negative control group	0	1	1	1	1	1	1
Three ST groups with different concentrations	2.5	1.06 ± 0.06	2.30 ± 0.93a	1.08 ± 0.04	1.10 ± 0.12	1.13 ± 0.11	3.09 ± 0.59a
	5	1.34 ± 0.25a	9.02 ± 0.32b	1.47 ± 0.31a	1.50 ± 0.27a	1.43 ± 0.16a	9.46 ± 0.58b
	10	1.63 ± 0.14a	31.34 ± 0.13b	2.02 ± 0.71a	1.82 ± 0.40a	1.75 ± 0.25a	47.29 ± 0.68b
Positive control group	5	1.33 ± 0.17a	10.02 ± 0.57b	1.49 ± 0.30a	1.50 ± 0.19a	1.47 ± 0.11a	8.92 ± 0.09b

By comparing the ST group with the negative control group.

^aP < 0.05,

^bP < 0.01, vs control groups. ST: Solanum lyratum Thumb.